Ubiquitinated Proteomic Study of Muscle Energy Metabolism of FAD3B Transgenic Mice

doi:10.7668/hbnxb.20195968

Abstract

Abstract:

The FAD3 gene in plants plays a crucial role in regulating polyunsaturated fatty acid conversion and energy metabolism.To investigate the impact of FAD3B on muscle energy metabolism in mice,leg muscles were isolated from FAD3B transgenic mice (MF) and wild-type mice (MW),followed by total protein extraction.Protein samples were qualitatively and quantitatively analyzed,and peptides were separated via LC-MS/MS using a strong cation exchange column to obtain raw mass spectrometry data.The data were processed using mass spectrometry software,and a peptide-protein dataset with significant peaks was selected for cluster heatmap visualization to compare MF and MW protein samples.Subsequently,biostatistical and bioinformatics analyses,including functional prediction,were performed to identify candidate target proteins regulated by FAD3B that were ubiquitinated and associated with muscle energy metabolism.The results revealed 180 differentially ubiquitinated peptides enriched in 65 proteins.Among these,seven upregulated peptides were enriched in seven proteins,while 54 downregulated peptides were enriched in 31 proteins.GO analysis,KEGG analysis and STRING analysis were performed on all target proteins enriched with differentially ubiquitinated peptides,and it was found that most of the target proteins were related to glycolysis,muscle contraction,protein synthesis and catabolism.In conclusion,exogenous FAD3B gene transfer alters ubiquitination modification levels in cellular structural components,muscle development,and energy metabolism-related proteins in mice.This demonstrates that FAD3B may influence skeletal muscle growth not only by regulating polyunsaturated fatty acid conversion but also by modulating ubiquitination of key proteins.These results provide a novel theoretical foundation and molecular targets for studying the mechanism of FAD3B in skeletal muscle development.

Key words: Mice, FAD3 gene, Ubiquitinated proteomics, Ubiquitination, Bioinformatics analysis, Muscle energy metabolism

CLC Number:

S813.3生物技术遗传育种

DU Xinyu, GUO Yunpeng, CHEN Zhuo, YANG Xinyue, LI Xin, JU Jiawen, GUO Yiwen. Ubiquitinated Proteomic Study of Muscle Energy Metabolism of FAD3B Transgenic Mice[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(6): 223-230. doi: 10.7668/hbnxb.20195968.

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Figures/Tables 5

Fig.1 Statistical analysis of ubiquitinated proteomics identification results A.Coomassie brilliant blue stain;B.Heat map of MF and MW ubiquitinated proteins clustering.

Tab.1 Protein enrichment table for some differential peptides

上调差异肽段富集情况 Up-regulated differential peptide enrichment		下调差异肽段富集情况 Down-regulated differential peptide enrichment
蛋白名 Protein name	蛋白功能描述 Protein function description	蛋白名 Protein name	蛋白功能描述 Protein function description
Ttn	肌联蛋白	Mylpf	肌球蛋白调节轻链 11
Neb	星云蛋白	Tpm1	原肌球蛋白 α-1 链
Eef2	真核翻译延伸因子2	Cul5	泛素连接酶
Ube2m	泛素结合酶E2M	Tnni2	肌钙蛋白 I,快速骨骼肌
Acta1	肌动蛋白,α 骨骼肌	Des	结蛋白
Mybpc2	肌球蛋白结合蛋白 C,快型	Gapdh	甘油醛-3-磷酸脱氢酶
Tmem30a	跨膜蛋白30a	Uba1	泛素样修饰剂激活酶 1
		Nedd8	泛素样蛋白
		Itm2c	整合膜蛋白 2C
		Myoz1	肌增生素1

Fig.2 GO enrichment analysis of differentially expressed ubiquitinated proteins 1.Canonical glycolysis;2.Muscle contraction;3.Sarcomere organization;4.Striated muscle contraction;5.Glycolytic process;6.Regulation of muscle contraction;7.Cardiac muscle contraction;8.Gluconeogenesis;9.Ubiquitin-dependent protein catabolic process;10.Protein ubiquitination;11.Striated muscle thin filament;12.M band;13.Sarcomere;14.Myofibril;15.Myosin complex;16.Cytosolic small ribosomal subunit;17.Z disc;18.Myelin sheath;19.Macromolecular complex;20.Cytosol;21.Oxygen transporter activity;22.Structural constituent of muscle;23.Oxygen binding;24.Structural constituent of cytoskeleton;25.Ubiquitin binding;26.Actin filament binding;27.Ion channel binding;28.Actin binding;29.Macromolecular complex binding;30.ATP binding.

Fig.3 KEGG enrichment analysis of differentially expressed ubiquitinated proteins 1.Motor proteins;2.Glycolysis/Gluconeogenesis;3.Hypertrophic cardiomyopathy;4.Dilated cardiomyopathy;5.Ubiquitin mediated proteolysis;6.Adrenergic signaling in cardiomyocytes;7.Biosynthesis of amino acids;8.Malaria;9.Carbon metabolism;10.African trypanosomiasis;11.HIF-1 signaling pathway;12.Arrhythmogenic right ventricular cardiomyopathy;13.Cardiac muscle contraction;14.Necroptosis;15.Parkinson disease;16.Pathway of neurodegeneration-multiple diseases;17.Oocyte meiosis;18.Nucleotide excision repair;19.Calcium signaling pathway.

Fig.4 STRING analysis of differentially ubiquitinated proteins

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